THE VP8 FRAGMENT OF VP4 IS THE RHESUS ROTAVIRUS HEMAGGLUTININ
THE VP8 FRAGMENT OF VP4 IS THE RHESUS ROTAVIRUS HEMAGGLUTININ
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DOI：
10.1016/0042-6822(91)90888-i
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发表时间：
1991-04-01
期刊：
影响因子：
3.7
通讯作者：
MACKOW, ER
中科院分区：
文献类型：
--
作者：
FIORE, L;GREENBERG, HB;MACKOW, ER
用重组杆状病毒在SF-9细胞中表达了VP4的氨基末端胰酶裂解片段VP8。杆状病毒表达的VP8蛋白是抗原性保守的，通过中和单抗文库的识别证明了这一点。在SF-9细胞中，表达的VP8蛋白能够凝集人0型红细胞，表明247个氨基酸的VP8胰酶切割片段中含有功能完整的恒河猴轮状病毒血凝素。VP8与呼肠孤病毒α-1蛋白的氨基酸末端282个氨基酸的相似性表明，α-1的血凝功能也存在于这些氨基酸末端。用表达的VP8蛋白免疫小鼠，获得了广泛交叉反应的中和抗体反应。表达的VP8蛋白的抗体可中和血清1-4和6的病毒，但不能中和猪OSU(ST5)或Gottfred(ST4)株。对VP8的中和抗体反应似乎比对表达的VP4或对整个RRV病毒粒子的免疫反应更具交叉反应。这表明亚单位蛋白免疫可以扩大对轮状病毒的中和抗体免疫应答，并增强对不同血清型毒株的保护性免疫。
The amino-terminal trypsin cleavage fragment of VP4, called VP8, was expressed from a recombinant baculovirus in Sf-9 cells. The baculovirus-expressed VP8 protein is antigenically conserved as demonstrated by its recognition by a library of neutralizing monoclonal antibodies. In Sf-9 cell sonicates, the expressed VP8 protein is capable of agglutinating human type 0 erythrocytes, indicating that the functionally intact rhesus rotavirus viral hemagglutinin is contained in the 247-amino acid VP8 trypsin cleavage fragment. Amino acid similarities between VP8 and the amino-terminal 282 amino acids of the reovirus α1 protein suggests that the α1 hemagglutination function resides within these amino-terminal amino acids as well. When the expressed VP8 protein was used to immunize mice, a broadly cross-reactive neutralizing antibody response was obtained. Antibodies elicited to the expressed VP8 protein neutralized viruses of serotypes 1–4 and 6 but not porcine strains OSU (st5) or Gottfried (st4). The neutralizing antibody response to VP8 appeared to be more cross-reactive than the immune response to expressed VP4 or to whole RRV virion. This suggests that subunit protein immunizations may broaden the neutralizing antibody immune responses to rotaviruses and enhance protective immunity to serotypically distinct strains.